Delay of postharvest ripening and senescence of tomato fruit through virus-induced LeACS2 gene silencing

Plant virus-induced gene silencing (VIGS) is currently a powerful tool for the study of gene function in plants. Here we report the silencing of LeACS2 by vacuum-infiltration and the tobacco rattle virus (TRV)-based VIGS method, which leads to a significant delay of the postharvest ripening and senescence of tomato fruit. Harvested mature green tomato fruit were vacuum-infiltrated with Agrobacterium strain GV3101 containing pTRV1 and pTRV2-LeACS2. Because of the silencing of LeACS2, the ethylene climacteric and pigment changes were clearly delayed. The onset of fruit ripening and senescence was significantly postponed, and transcription of LeACS2 and ACC synthase activity were also suppressed in treated tomato fruit during storage. The silencing of LeACS2 by vacuum infiltration, however, did not alter the contents assayed at the end of storage. Our results indicate that vacuum infiltration is a highly efficient TRV-based VIGS method to silence LeACS2 in harvested tomato fruit. It can obviously delay ripening and senescence, and is a potential method for postharvest preservation of tomato fruit.

[1]  T. Goodwin.  Chemistry and biochemistry of plant pigments , 1976 .

[2]  A. Kander.  Composition and flavor quality of fresh market tomatoes as influenced by some postharvest handling procedures , 1978 .

[3]  R. Buttery,et al.  Effect of storage and ripening on fresh tomato quality, Part I , 1994 .

[4]  D. Grierson,et al.  Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening. , 2002, Journal of experimental botany.

[5]  D. Olson,et al.  Differential expression of two genes for 1-aminocyclopropane-1-carboxylate synthase in tomato fruits. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[6]  M. Tomes.  Temperature Inhibition of Carotene Synthesis in Tomato , 1963, Botanical Gazette.

[7]  T. Zarembinski,et al.  Ethylene biosynthesis and action: a case of conservation , 1994, Plant Molecular Biology.

[8]  K. Gross.  A rapid and sensitive spectrophotometric method for assaying polygalacturonase using 2-cyanoacetamide [Tomato, fruit softening] , 1982 .

[9]  Kun Luo,et al.  Enhancement of virus-induced gene silencing in tomato by low temperature and low humidity. , 2006, Molecules and cells.

[10]  D. Robertson.  VIGS vectors for gene silencing: many targets, many tools. , 2004, Annual review of plant biology.

[11]  S. Shiomi,et al.  Differential expression and internal feedback regulation of 1-aminocyclopropane-1-carboxylate synthase, 1-aminocyclopropane-1-carboxylate oxidase, and ethylene receptor genes in tomato fruit during development and ripening. , 1998, Plant physiology.

[12]  J. Fellman,et al.  A role for jasmonates in climacteric fruit ripening , 1998, Planta.

[13]  D. Olson,et al.  Differential induction of seven 1-aminocyclopropane-1-carboxylate synthase genes by elicitor in suspension cultures of tomato (Lycopersicon esculentum) , 1997, Plant Molecular Biology.

[14]  C. Wang,et al.  Modified atmosphere packaging of fruits and vegetables. , 1989, Critical reviews in food science and nutrition.

[15]  M. Taylor,et al.  Potato Virus X-Induced Gene Silencing in Leaves and Tubers of Potato1 , 2004, Plant Physiology.

[16]  Hongliang Zhu,et al.  Virus-induced gene silencing in tomato fruit. , 2005, The Plant journal : for cell and molecular biology.

[17]  G. Barry,et al.  Control of ethylene synthesis by expression of a bacterial enzyme in transgenic tomato plants. , 1991, The Plant cell.

[18]  J. Kirk.  Studies on the dependence of chlorophyll synthesis on protein synthesis in Euglena gracilis, together with a nomogram for determination of chlorophyll concentration , 1967, Planta.

[19]  D. Grierson,et al.  Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants , 1990, Nature.

[20]  Shangfa Yang,et al.  A simple and sensitive assay for 1-aminocyclopropane-1-carboxylic acid. , 1979, Analytical biochemistry.

[21]  M. Redinbaugh,et al.  A procedure for the small-scale isolation of plant RNA suitable for RNA blot analysis. , 1988, Analytical biochemistry.

[22]  P. Oeller,et al.  LE-ACS4, a fruit ripening and wound-induced 1-aminocyclopropane-1-carboxylate synthase gene of tomato (Lycopersicon esculentum). Expression in Escherichia coli, structural characterization, expression characteristics, and phylogenetic analysis. , 1993, The Journal of biological chemistry.

[23]  D. Baulcombe.  RNA silencing in plants , 2004, Nature.

[24]  P. Oeller,et al.  Reversible inhibition of tomato fruit senescence by antisense RNA. , 1991, Science.

[25]  A. Halevy,et al.  The mechanism involved in ethylene-enhanced ethylene synthesis in carnations , 1994, Plant growth regulation (Print).

[26]  S. Dinesh-Kumar,et al.  Virus-induced gene silencing. , 2003, Methods in molecular biology.

[27]  S. Dinesh-Kumar,et al.  Virus-induced gene silencing in tomato. , 2002, The Plant journal : for cell and molecular biology.

[28]  A. Bennett,et al.  Polygalacturonase Gene Expression in Rutgers, rin, nor, and Nr Tomato Fruits. , 1987, Plant physiology.